Objective: Pseudomonas aeruginosa is an opportunistic pathogen that infects hospitalized, burned and immunosuppressed patients. The main aim of the present study was to develop a vaccination strategy based on recombinant flagellin type A (r-fla-A) that would enhance the protective response against P. aeruginosa in the burn wound sepsis model. Materials and Methods: After the preparation of type A r-flagellin, a specific polyclonal IgG was produced in rabbit. After immediate post-burn and post-challenge, mortality rate was screened in the mice treated with anti r-fla-A IgG, and inbred mice were also immunized with r-fla-A in separate groups. After final booster, vaccinated mice were burned and challenged with P. aeruginosa. The functional activity of anti r-flagellin antisera was determined by opsonophagocytic killing test. To evaluate the humoral immune response, sera were analyzed by ELISA for its total antibody. Results: In vivo administration of r-fla-A afforded a remarkable improvement in the survival of mice challenged with homologous strain (PAK) in the burn wound infection (83.3% vs. 0% in control; P < 0.005). The antibodies generated against the r-fla-A achieved 25% survival in immunized mice that were infected with heterologous strain PAO1. The anti r-fla-A IgG afforded a significant improvement in survival of mice infected by homologous strain PAK from 16.6% to 75%; In contrast, this antiserum achieved 33.3% survival following challenge with heterologous strain PAO1 (compared to control IgG). Anti r-flagellin antibody promoted phagocytosis of the homologous strain and decreased the killing of heterologous strain (53.1% and 17.4% respectively vs. 3.7% in the control group; P < 0.001). Flagellin also induced a high level humoral immune response in the immunized burned and challenged mice. Conclusion: We concluded that immunization with r-fla-A and anti r-fla-A would protect burned mice against lethal P. aeruginosa challenge.